Journal List > Korean J Obstet Gynecol > v.53(1) > 1006402

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Park, Park, Kim, Ro, Kim, Jung, Bae, and Ahn: Comparison of cell growth suppression in SiHa cervical carcinoma cell line by human papillomavirus type 16 E6/E7 siRNAs
Original Article
Korean Journal of Obstetrics and Gynecology

Korean Journal of Obstetrics and Gynecology 2010; 53(1): 35-42.

Published online: 31 January 2010

DOI: https://doi.org/10.5468/kjog.2010.53.1.35

Comparison of cell growth suppression in SiHa cervical carcinoma cell line by human papillomavirus type 16 E6/E7 siRNAs

Sae-Hyun Park, M.D.1, Byung-Joon Park, M.D.1, Yong-Wook Kim, M.D.1, Duck-Yeong Ro, M.D.1, Tae-Eung Kim, M.D.1, Jae-Keun Jung, M.D.1, Su-Mi Bae, Ph.D.2, Woong-Shick Ahn, M.D., Ph.D.1

1Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea, Seoul, Korea.

2Department of Catholic Research Institutes of Medical Science, College of Medicine, The Catholic University of Korea, Seoul, Korea.

Corresponding author (ahnlab1@catholic.ac.kr)

Received 31 May 2008       Accepted 16 November 2009

Copyright © 2010 Korean Society of Obstetrics and Gynecology

Abstract

Objective

Human cervical cancer is caused by the high-risk types of human papillomavirus (HPV) such as HPV16, which possess the E6 and E7 oncogenes, whose expressions are a prerequisite for cancer development. We performed this study to compare the efficacy of antitumor activity by HPV siRNA which silences only E6 or both E6/E7.

Methods

We transfected siRNA 377 (HPV16 E6 siRNA), siRNA 3 (HPV16 E6 siRNA), and siRNA 198 (HPV16 E7 siRNA) into SiHa cell line (siRNA 377 silences only E6, and siRNA 3 and siRNA 198 silence both E6 and E7). We experimented cell counts and morphologic changes 24 and 48 hours after transfection and expressions of HPV16 E6/E7 mRNA by RT-PCR.

Results

siRNA 377, siRNA 3, and siRNA 198 suppressed the cell growth. siRNA 3 and siRNA 198 were more potent than siRNA 377 in cell growth suppression. siRNA 377 knocked down the expression of E6 mRNA, and both siRNA 3 and siRNA 198 knocked down the expression of E6/E7 mRNA.

Conclusion

Our results suggest that simultaneous suppression of E6 and E7 was more potent than E6-specific suppression in cancer cell growth.

Keywords: Cervical cancer, Human papillomavirus, E6, E7, siRNA

Figures and Tables

Fig. 1
Effects of siRNA on growth inhibition in SiHa cervical cancer cell line in vitro. Cells were transfected with universal siRNA, siRNA 3, siRNA 198 and siRNA 377 at 50 nM. Cells were trypsinized for direct cell counting after 24 and 48 hours. Cell growth inhibitions were compared with the universal siRNA control by t-test. *P<0.05, **P<0.01.
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Fig. 2
siRNA-mediated morphological and numerical changes in SiHa cervical cancer cell line in vitro. Cells were transfected with universal siRNA, siRNA 3, siRNA 198 and siRNA 377 at 50 nM. Typical photographs were shown after 48 hours (×400 for all photographs).
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Fig. 3
RT-PCR analysis for expressions of the HPV16 E6, E7 and β-actin mRNA. Cells were transfected with universal siRNA, siRNA 377, siRNA 3 and siRNA 198 at 50 and 100 nM. RNA was extracted for RT-PCR after 48 hours.
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Fig. 4
Effects of siRNA on p53 expressions in SiHa cell line. Cells were transfected with universal siRNA, siRNA 377, siRNA 3 and siRNA 198 at 50 nM. Immunoblot analysis using specific p53 antibody was performed after 24 and 48 hours. The cells were collected and the extracts (40 µg) were run on 10% SDS-PAGE.
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Table 1
Oligomer sequences used for gene silencing
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Table 2
Primer sequences used for RT-PCR
kjog-53-35-i002

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